Standardized bacteriophage purification for personalized phage therapy

Abstract

The world is on the cusp of a post-antibiotic era, but researchers and medical doctors have found a way forward—by looking back at how infections were treated before the advent of antibiotics, namely using phage therapy. Although bacteriophages (phages) continue to lack drug approval in Western medicine, an increasing number of patients are being treated on an expanded-access emergency investigational new drug basis. To streamline the production of high-quality and clinically safe phage preparations, we developed a systematic procedure for medicinal phage isolation, liter-scale cultivation, concentration and purification. The 16- to 21-day procedure described in this protocol uses a combination of modified classic techniques, modern membrane filtration processes and no organic solvents to yield on average 23 mL of 1011 plaque-forming units (PFUs) per milliliter for Pseudomonas, Klebsiella, and Serratia phages tested. Thus, a single production run can produce up to 64,000 treatment doses at 109 PFUs, which would be sufficient for most expanded-access phage therapy cases and potentially for clinical phase I/II applications. The protocol focuses on removing endotoxins early by conducting multiple low-speed centrifugations, microfiltration, and cross-flow ultrafiltration, which reduced endotoxins by up to 106-fold in phage preparations. Implementation of a standardized phage cultivation and purification across research laboratories participating in phage production for expanded-access phage therapy might be pivotal to reintroduce phage therapy to Western medicine.

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Fig. 1: Overview of bacteriophage cultivation and purification.
Fig. 2: Schematic of phage lysate dead-end filtration (Step 91) and cross-flow filtration (Steps 93–104) removal of impurities.
Fig. 3: Process stepwise phage titer and endotoxin concentration throughout processing.
Fig. 4: Purity and safety analyses of final phage preparations.

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

References

  1. 1.

    Schooley, R. T. et al. Development and use of personalized bacteriophage-based therapeutic cocktails to treat a patient with a disseminated resistant Acinetobacter baumannii infection. Antimicrob. Agents Chemother. 61, e00954–17 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Dedrick, R. M. et al. Engineered bacteriophages for treatment of a patient with a disseminated drug-resistant Mycobacterium abscessus. Nat. Med. 25, 730–733 (2019).

    CAS  PubMed  PubMed Central  Google Scholar 

  3. 3.

    Onsea, J. et al. Bacteriophage application for difficult-to-treat musculoskeletal infections: development of a standardized multidisciplinary treatment protocol. Viruses 11, 891 (2019).

  4. 4.

    Nir-Paz, R. et al. Successful treatment of antibiotic resistant poly-microbial bone infection with bacteriophages and antibiotics combination. Clin. Infect. Dis. 69, 2015–2018 (2019).

  5. 5.

    Van Norman, G. A. Expanding patient access to investigational drugs: single patient investigational new drug and the “right to try”. JACC Basic Transl. Sci. 3, 280–293 (2018).

    PubMed  PubMed Central  Google Scholar 

  6. 6.

    Svircev, A., Roach, D. & Castle, A. Framing the future with bacteriophages in agriculture. Viruses 10, 218 (2018).

  7. 7.

    Segall, A. M., Roach, D. R. & Strathdee, S. A. Stronger together? Perspectives on phage–antibiotic synergy in clinical applications of phage therapy. Curr. Opin. Microbiol. 51, 46–50 (2019).

    PubMed  Google Scholar 

  8. 8.

    Moye, Z. D., Woolston, J. & Sulakvelidze, A. Bacteriophage applications for food production and processing. Viruses 10, 205 (2018).

    PubMed Central  Google Scholar 

  9. 9.

    Lehman, S. M. et al. Design and preclinical development of a phage product for the treatment of antibiotic-resistant Staphylococcus aureus infections. Viruses 11, 88 (2019).

    CAS  PubMed Central  Google Scholar 

  10. 10.

    Pirnay, J.-P. et al. Quality and safety requirements for sustainable phage therapy products. Pharm. Res. 32, 2173–2179 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Mutti, M. & Corsini, L. Robust approaches for the production of active ingredient and drug product for human phage therapy. Front. Microbiol. 10, 2289 (2019).

  12. 12.

    Kutateladze, M. & Adamia, R. Phage therapy experience at the Eliava Institute. Med. Mal. Infect. 38, 426–430 (2008).

    CAS  PubMed  Google Scholar 

  13. 13.

    Letkiewicz, S. et al. The perspectives of the application of phage therapy in chronic bacterial prostatitis. FEMS Immunol. Med. Microbiol. 60, 99–112 (2010).

    CAS  PubMed  Google Scholar 

  14. 14.

    Jault, P. et al. Efficacy and tolerability of a cocktail of bacteriophages to treat burn wounds infected by Pseudomonas aeruginosa (PhagoBurn): a randomised, controlled, double-blind phase 1/2 trial. Lancet Infect. Dis. 19, 35–45 (2019).

    PubMed  Google Scholar 

  15. 15.

    US Department of Health and Human Services (Antibiotic Resistance Coordination and Strategy Unit within the Division of Healthcare Quality Promotion, Centers for Disease Control and Prevention, 2019).

  16. 16.

    O’Neill, J. Tackling Drug-Resistant Infections Globally: Final Report and Recommendations (Department of Health, United Kingdom, 2016).

  17. 17.

    d’Hérelle, F. L'étude d’une maladie: le choléra, maladie à paradoxes (Rouge, 1946).

  18. 18.

    Adams, M. Enumeration of bacteriophage particles. In: Bacteriophages (ed Adams MH) 27–34 (Interscience Publishers, 1959).

  19. 19.

    Roach, D. R. & Debarbieux, L. Phage therapy: awakening a sleeping giant. Emerg. Top. Life Sci. 1, 93–103 (2017).

    CAS  PubMed Central  Google Scholar 

  20. 20.

    Dalpke, A., Frank, J., Peter, M. & Heeg, K. Activation of toll-like receptor 9 by DNA from different bacterial species. Infect. Immun. 74, 940–946 (2006).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. 21.

    Sweere, J. M. et al. Bacteriophage trigger antiviral immunity and prevent clearance of bacterial infection. Science 363, eaat9691 (2019).

    CAS  PubMed  PubMed Central  Google Scholar 

  22. 22.

    Opal, S. M. Endotoxins and other sepsis triggers. Contrib. Nephrol. 167, 14–24 (2010).

    CAS  PubMed  Google Scholar 

  23. 23.

    Capparelli, R., Parlato, M., Borriello, G., Salvatore, P. & Iannelli, D. Experimental phage therapy against Staphylococcus aureus in mice. Antimicrob. Agents Chemother. 51, 2765 (2007).

    CAS  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Forti, F. et al. Design of a broad-range bacteriophage cocktail that reduces Pseudomonas aeruginosa biofilms and treats acute infections in two animal models. Antimicrob. Agents Chemother. 62, e02573–17 (2018).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Roach, D. R. et al. Synergy between the host immune system and bacteriophage is essential for successful phage therapy against an acute respiratory pathogen. Cell Host Microbe 22, 38–47 (2017).

    CAS  PubMed  Google Scholar 

  26. 26.

    Chibani-Chennoufi, S. et al. In vitro and in vivo bacteriolytic activities of Escherichia coli phages: implications for phage therapy. Antimicrob. Agents Chemother. 48, 2558 (2004).

    CAS  PubMed  PubMed Central  Google Scholar 

  27. 27.

    Yamamoto, K. R., Alberts, B. M., Benzinger, R., Lawhorne, L. & Treiber, G. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology 40, 734–744 (1970).

    CAS  PubMed  Google Scholar 

  28. 28.

    Bachrach, U. & Friedmann, A. Practical procedures for the purification of bacterial viruses. Appl. Microbiol. 22, 706–715 (1971).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. 29.

    Ackermann, H. W. et al. Guidelines for bacteriophage characterization. Adv. Virus Res. 23, 1–24 (1978).

    CAS  PubMed  Google Scholar 

  30. 30.

    Boratynski, J. et al. Preparation of endotoxin-free bacteriophages. Cell Mol. Biol. Lett. 9, 253–259 (2004).

    CAS  PubMed  Google Scholar 

  31. 31.

    Gill, J. & Hyman, P. Phage choice, isolation, and preparation for phage therapy. Curr. Pharm. Biotechnol. 11, 2–14 (2010).

    CAS  PubMed  Google Scholar 

  32. 32.

    Bourdin, G. et al. Amplification and purification of T4-like Escherichia coli phages for phage therapy: from laboratory to pilot scale. Appl. Environ. Microbiol. 80, 1469–1476 (2014).

    PubMed  PubMed Central  Google Scholar 

  33. 33.

    Bonilla, N. et al. Phage on tap-a quick and efficient protocol for the preparation of bacteriophage laboratory stocks. PeerJ 4, e2261 (2016).

    PubMed  PubMed Central  Google Scholar 

  34. 34.

    Van Belleghem, J. D., Merabishvili, M., Vergauwen, B., Lavigne, R. & Vaneechoutte, M. A comparative study of different strategies for removal of endotoxins from bacteriophage preparations. J. Microbiol. Methods 132, 153–159 (2017).

    PubMed  Google Scholar 

  35. 35.

    Van Belleghem, J. D., Dabrowska, K., Vaneechoutte, M., Barr, J. J. & Bollyky, P. L. Interactions between bacteriophage, bacteria, and the mammalian immune system. Viruses 11, 10 (2018).

  36. 36.

    Gogokhia, L. et al. Expansion of bacteriophages is linked to aggravated intestinal inflammation and colitis. Cell Host Microbe 25, 285–299 (2019).

    CAS  PubMed  PubMed Central  Google Scholar 

  37. 37.

    An, T. W., Kim, S. J., Lee, Y. D., Park, J. H. & Chang, H. I. The immune-enhancing effect of the Cronobacter sakazakii ES2 phage results in the activation of nuclear factor-kappaB and dendritic cell maturation via the activation of IL-12p40 in the mouse bone marrow. Immunol. Lett. 157, 1–8 (2014).

    CAS  PubMed  Google Scholar 

  38. 38.

    Van Belleghem, J. D., Clement, F., Merabishvili, M., Lavigne, R. & Vaneechoutte, M. Pro- and anti-inflammatory responses of peripheral blood mononuclear cells induced by Staphylococcus aureus and Pseudomonas aeruginosa phages. Sci. Rep. 7, 8004 (2017).

    PubMed  PubMed Central  Google Scholar 

  39. 39.

    Chan, B. K. et al. Phage treatment of an aortic graft infected with Pseudomonas aeruginosa. Evol. Med. Public Health 2018, 60–66 (2018).

    PubMed  PubMed Central  Google Scholar 

  40. 40.

    Pirnay, J. P. et al. The magistral phage. Viruses 10, 64 (2018).

  41. 41.

    US Department of Health and Human Services Food and Drug Administration (Office of Communications, Division of Drug Information (US Food and Drug Administration, 2012).

  42. 42.

    Spaulding, A. R. et al. Staphylococcal and streptococcal superantigen exotoxins. Clin. Microbiol. Rev. 26, 422–447 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  43. 43.

    Carlson, K. Working with Bacteriophages: Common Techniques and Methodological Approaches, Vol 1 (CRC Press, 2005).

  44. 44.

    Davidson, I. W., Sumner, D. D. & Parker, J. C. Chloroform: a review of its metabolism, teratogenic, mutagenic, and carcinogenic potential. Drug Chem. Toxicol. 5, 1–87 (1982).

    CAS  PubMed  Google Scholar 

  45. 45.

    Szermer-Olearnik, B. & Boratyński, J. Removal of endotoxins from bacteriophage preparations by extraction with organic solvents. PLoS ONE 10, e0122672 (2015).

    PubMed  PubMed Central  Google Scholar 

  46. 46.

    Melnikov, P. & Zanoni, L. Z. Clinical effects of cesium intake. Biol. Trace Elem. Res. 135, 1–9 (2010).

    CAS  PubMed  Google Scholar 

  47. 47.

    Centeno, J. A. et al. Blood and tissue concentration of cesium after exposure to cesium chloride: a report of two cases. Biol. Trace Elem. Res. 94, 97–104 (2003).

    CAS  PubMed  Google Scholar 

  48. 48.

    Mancuso, F., Shi, J. & Malik, D. J. High throughput manufacturing of bacteriophages using continuous stirred tank bioreactors connected in series to ensure optimum host bacteria physiology for phage production. Viruses 10, 537 (2018).

    PubMed Central  Google Scholar 

  49. 49.

    Edelman, D. C. & Barletta, J. Real-time PCR provides improved detection and titer determination of bacteriophage. Biotechniques 35, 368–375 (2003).

    CAS  PubMed  Google Scholar 

  50. 50.

    Anderson, B. et al. Enumeration of bacteriophage particles: comparative analysis of the traditional plaque assay and real-time QPCR- and nanosight-based assays. Bacteriophage 1, 86–93 (2011).

    PubMed  PubMed Central  Google Scholar 

  51. 51.

    Freeman, V. J. Studies on the virulence of bacteriophage-infected strains of Corynebacterium diphtheriae. J. Bacteriol. 61, 675–688 (1951).

    CAS  PubMed  PubMed Central  Google Scholar 

  52. 52.

    Waldor, M. K. & Mekalanos, J. J. Lysogenic conversion by a filamentous phage encoding cholera toxin. Science 272, 1910–1914 (1996).

    CAS  PubMed  Google Scholar 

  53. 53.

    Hatano, M., Nakamura, K. & Kurokawa, M. Isolation of a new temperature phage causing the lysogenic conversion in Corynebacterium diphtheriae. Jpn. J. Microbiol. 3, 301–311 (1959).

    CAS  PubMed  Google Scholar 

  54. 54.

    Weeks, C. R. & Ferretti, J. J. The gene for type-A streptococcal exotoxin (erythrogenic toxin) is located in bacteriophage-T12. Infect. Immun. 46, 531–536 (1984).

    CAS  PubMed  PubMed Central  Google Scholar 

  55. 55.

    Schmidt, H. Shiga-toxin-converting bacteriophages. Res. Microbiol. 152, 687–695 (2001).

    CAS  PubMed  Google Scholar 

  56. 56.

    Plunkett, G., Rose, D. J., Durfee, T. J. & Blattner, F. R. Sequence of shiga toxin 2 phage 933W from Escherichia coli O157:H7: shiga toxin as a phage late-gene product. J. Bacteriol. 181, 1767–1778 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  57. 57.

    Cobian Guemes, A. G. et al. Cystic fibrosis rapid response: translating multi-omics data into clinically relevant information. mBio 10, e00431-19 (2019).

  58. 58.

    Eklund, M. W., Poysky, F. T., Reed, S. M. & Smith, C. A. Bacteriophage and the toxigenicity of Clostridium botulinum type C. Science 172, 480–482 (1971).

    CAS  PubMed  Google Scholar 

  59. 59.

    Brussow, H., Canchaya, C. & Hardt, W. D. Phages and the evolution of bacterial pathogens: from genomic rearrangements to lysogenic conversion. Microbiol. Mol. Biol. Rev. 68, 560–602 (2004).

    PubMed  PubMed Central  Google Scholar 

  60. 60.

    Wagner, P. L. & Waldor, M. K. Bacteriophage control of bacterial virulence. Infect. Immun. 70, 3985–3993 (2002).

    CAS  PubMed  PubMed Central  Google Scholar 

  61. 61.

    Brown-Jaque, M. et al. Antibiotic resistance genes in phage particles isolated from human faeces and induced from clinical bacterial isolates. Int. J. Antimicrob. Agents 51, 434–442 (2018).

    CAS  PubMed  Google Scholar 

  62. 62.

    Purdy, A., Rohwer, F., Edwards, R., Azam, F. & Bartlett, D. H. A glimpse into the expanded genome content of Vibrio cholerae through identification of genes present in environmental strains. J. Bacteriol. 187, 2992–3001 (2005).

    CAS  PubMed  PubMed Central  Google Scholar 

  63. 63.

    Enault, F. et al. Phages rarely encode antibiotic resistance genes: a cautionary tale for virome analyses. ISME J. 11, 237–247 (2017).

    CAS  PubMed  Google Scholar 

  64. 64.

    Billard-Pomares, T. et al. Characterization of a P1-like bacteriophage carrying an SHV-2 extended-spectrum beta-lactamase from an Escherichia coli strain. Antimicrob. Agents Chemother. 58, 6550–6557 (2014).

    PubMed  PubMed Central  Google Scholar 

  65. 65.

    Zhou, W., Liu, L., Feng, Y. & Zong, Z. A P7 phage-like plasmid carrying mcr-1 in an ST15 Klebsiella pneumoniae clinical isolate. Front. Microbiol. 9, 11 (2018).

    PubMed  PubMed Central  Google Scholar 

  66. 66.

    Aziz, R. K. et al. Mosaic prophages with horizontally acquired genes account for the emergence and diversification of the globally disseminated M1T1 clone of Streptococcus pyogenes. J. Bacteriol. 187, 3311–3318 (2005).

    CAS  PubMed  PubMed Central  Google Scholar 

  67. 67.

    Lacey, J. A., Johanesen, P. A., Lyras, D. & Moore, R. J. In silico identification of novel toxin homologs and associated mobile genetic elements in Clostridium perfringens. Pathogens 8 (2019).

  68. 68.

    Nguyen, M. et al. Using machine learning to predict antimicrobial MICs and associated genomic features for nontyphoidal Salmonella. J. Clin. Microbiol. 57, e1260-18 (2019).

  69. 69.

    Jia, B. et al. CARD 2017: expansion and model-centric curation of the comprehensive antibiotic resistance database. Nucleic Acids Res. 45, D566–D573 (2017).

    CAS  PubMed  Google Scholar 

  70. 70.

    Zankari, E. et al. Identification of acquired antimicrobial resistance genes. J. Antimicrob. Chemother. 67, 2640–2644 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  71. 71.

    Feldgarden, M. et al. Validating the AMRFinder Tool and Resistance Gene Database by using antimicrobial resistance genotype-phenotype correlations in a collection of isolates. Antimicrob. Agents Chemother. 63, e00483–19 (2019).

    CAS  PubMed  PubMed Central  Google Scholar 

  72. 72.

    Gupta, S. K. et al. ARG-ANNOT, a new bioinformatic tool to discover antibiotic resistance genes in bacterial genomes. Antimicrob. Agents Chemother. 58, 212–220 (2014).

    PubMed  PubMed Central  Google Scholar 

  73. 73.

    Chen, L., Zheng, D., Liu, B., Yang, J. & Jin, Q. VFDB 2016: hierarchical and refined dataset for big data analysis-10 years on. Nucleic Acids Res. 44, D694–D697 (2016).

    CAS  PubMed  Google Scholar 

  74. 74.

    Carattoli, A. et al. In silico detection and typing of plasmids using PlasmidFinder and plasmid multilocus sequence typing. Antimicrob. Agents Chemother. 58, 3895–3903 (2014).

    PubMed  PubMed Central  Google Scholar 

  75. 75.

    Beaulaurier, J. et al. Assembly-free single-molecule sequencing recovers complete virus genomes from natural microbial communities. Genome Res. 30, 437–446 (2020).

    PubMed  PubMed Central  Google Scholar 

  76. 76.

    Wattam, A. R. et al. Improvements to PATRIC, the all-bacterial bioinformatics database and analysis resource center. Nucleic Acids Res. 45, D535–D542 (2017).

    CAS  PubMed  Google Scholar 

  77. 77.

    McNair, K., Bailey, B. A. & Edwards, R. A. PHACTS, a computational approach to classifying the lifestyle of phages. Bioinformatics 28, 614–618 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  78. 78.

    Edwards, R. A., McNair, K., Faust, K., Raes, J. & Dutilh, B. E. Computational approaches to predict bacteriophage–host relationships. FEMS Microbiol. Rev. 40, 258–272 (2016).

    CAS  PubMed  Google Scholar 

  79. 79.

    McNair, K. et al. in Bacteriophages: Methods and Protocols Vol 3 (eds Clokie, M.R.J., Kropinski, A.M. & Lavigne, R.) 231–238 (Springer, 2018).

  80. 80.

    Philipson, C. W. et al. Characterizing phage genomes for therapeutic applications. Viruses 10, 188 (2018).

  81. 81.

    Kutter, E. in Bacteriophages (eds Clokie, M.R.J. & Kropinski A.M.) 141–149 (Springer, 2009).

  82. 82.

    Erickson, H. P. Size and shape of protein molecules at the nanometer level determined by sedimentation, gel filtration, and electron microscopy. Biol. Proced. Online 11, 32 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  83. 83.

    Brakke, M. K. Density gradient centrifugation - a new separation technique. J. Am. Chem. Soc. 73, 1847–1848 (1951).

    CAS  Google Scholar 

  84. 84.

    Petsch, D. & Anspach, F. B. Endotoxin removal from protein solutions. J. Biotechnol. 76, 97–119 (2000).

    CAS  PubMed  Google Scholar 

  85. 85.

    Barr, J. J. et al. Bacteriophage adhering to mucus provide a non–host-derived immunity. Proc. Natl Acad. Sci. USA 110, 10771 (2013).

    CAS  PubMed  Google Scholar 

  86. 86.

    Bocian, K. et al. The effects of T4 and A3/R bacteriophages on differentiation of human myeloid dendritic cells. Front. Microbiol. 7 (2016).

  87. 87.

    Riss, T. L. et al. in Assay Guidance Manual (eds Sittampalam, G. S. et al.) (Eli Lilly & Company and the National Center for Advancing Translational Sciences, 2004).

  88. 88.

    Moir, D. T., Ming, D., Opperman, T., Schweizer, H. P. & Bowlin, T. L. A high-throughput, homogeneous, bioluminescent assay for Pseudomonas aeruginosa gyrase inhibitors and other DNA-damaging agents. J. Biomol. Screen. 12, 855–864 (2007).

    CAS  PubMed  PubMed Central  Google Scholar 

  89. 89.

    Van Twest, R. & Kropinski, A. M. Bacteriophage enrichment from water and soil. Methods Mol. Biol. 501, 15–21 (2009).

    PubMed  Google Scholar 

  90. 90.

    Summer, E. J. Preparation of a phage DNA fragment library for whole genome shotgun sequencing. Methods Mol. Biol. 502, 27–46 (2009).

    CAS  PubMed  Google Scholar 

  91. 91.

    Koren, S. et al. Canu: scalable and accurate long-read assembly via adaptive k-mer weighting and repeat separation. Genome Res. 27, 722–736 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  92. 92.

    Henry, M., Lavigne, R. & Debarbieux, L. Predicting in vivo efficacy of therapeutic bacteriophages used to treat pulmonary infections. Antimicrob. Agents Chemother. 57, 5961–5968 (2013).

    PubMed  PubMed Central  Google Scholar 

  93. 93.

    Boulanger, P. in Bacteriophages: Methods and Protocols Vol 2 (eds Clokie, M.R.J. & Kropinski A.M.) 227–238 (Humana Press, 2009).

  94. 94.

    Trend, S. et al. Use of a primary epithelial cell screening tool to investigate phage therapy in cystic fibrosis. Front. Pharmacol. 9, 1330 (2018).

    CAS  PubMed  PubMed Central  Google Scholar 

  95. 95.

    Roach, D. R., Sjaarda, D. R., Castle, A. J. & Svircev, A. M. Host exopolysaccharide quantity and composition impacts bacteriophage pathogenesis of Erwinia amylovora. Appl. Environ. Microbiol. 79, 3249–3256 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank J. Grose at Brigham Young University for gifting us phages JG265, JG266 and SM219 and the Serratia and Klebsiella strains. We thank R. Schooley at the University of California, San Diego for providing feedback. This research was supported in part by National Institutes of Health grant RC2DK116713 to R.A.E. and San Diego State University startup funds to D.R.R.

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D.R.R. and R.A.E. conceived the concepts and supervised the research and development. T.L. and A.C.S. conducted the in vitro experiments. R.A.E. performed the genomic analyses. All authors wrote and commented on the manuscript.

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Correspondence to Dwayne R. Roach.

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Key references using this protocol

Forti, F. et al. Antimicrob. Agents Chemother. 62, e02573-17 (2018): https://aac.asm.org/content/62/6/e02573-17

Roach, D. R. et al. Cell Host Microbe 22, 38–47 (2017): https://doi.org/10.1016/j.chom.2017.06.018

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Luong, T., Salabarria, A., Edwards, R.A. et al. Standardized bacteriophage purification for personalized phage therapy. Nat Protoc (2020). https://doi.org/10.1038/s41596-020-0346-0

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